On-Demand Fluorescent Reporter Assays

Features & Benefits

• High signal-to-noise ratio
• Bright signal
• Easy to monitor
• Red, green, and cyan options

Description

With the On-Demand Fluorescent Protein Reporter Systems, you can compensate for reporter background without lowering your assay’s signal intensity. Low background and a bright signal are no longer mutually exclusive.

The On-Demand Fluorescent Protein Reporter Systems meet the challenge by using a combination of technologies: each system includes a bright fluorescent protein reporter (AmCyan1, tdTomato, or ZsGreen1) for high signal intensity, coupled with ligand-dependent ProteoTuner™ protein stabilization/destabilization technology to eliminate background.

In these systems, the fluorescent protein reporter is expressed as a fusion protein tagged on its N-terminus with a ligand-dependent destabilization domain (DD). The DD rapidly targets the reporter protein for proteasomal degradation, guaranteeing a low reporter background signal at the start of your experiment. However, when the small, membrane-permeant ligand Shield1 is added to the sample, it binds to the DD and protects the reporter from degradation, so that it can accumulate.

By adding Shield1 simultaneously with your candidate inducer, you can effectively stabilize the reporter protein when it is synthesized in response to promoter activation. The majority of the fluorescent protein reporter molecules expressed during promoter activation will contribute to your readout, allowing for a considerably higher dynamic range and drastically improved signal-to-noise ratio compared to other types of reporter systems.